A new sulfonamide, 4-{(4-nitrophenylsulfonamido)methyl}cyclohexanecarboxylic acid (C14H18N2O6S), has been synthesized by the reaction of tranexamic acid and 4-nitroben-zenesulfonyl chloride in basic medium at room temperature. The molecular structure was determined by FT-IR, NMR, elemental analysis and single-crystal X-ray technique. X-ray diffraction shows that the compound crystallizes in the monoclinic system, space group P21/c with a = 13.5980(7), b = 4.9877(2), c = 23.3878(13) Å, β = 93.254(3)º, Z = 4, V = 1583.67(14) Å3, μ = 0.237 mm-1, F(000) = 720, R = 0.0471 and wR = 0.1182. The molecules are related by inversion and paired into dimers via C–H•••O interactions. The dimmers are interlinked due to strong N–H•••O bonds, where O-atoms are of sulfonyl groups. The molecules are stabilized in the form of infinite two-dimensional network with base vectors [0 1 0] and [0 0 –1] in the plane (1 0 2). The existence of good intermolecular interactions suggests the biological importance of the synthesized molecule. The compound was screened for its interaction with FS-DNA using UV-visible spectroscopy. UV-visible spectroscopic results depict that the compound interacts with DNA by mixed binding mode intercalation along with hydrogen bonding. Negative values of ΔG (–23.34, –17.79 kJ•mol−1) indicate spontaneity of the compound-DNA adduct formation.